1. Field of Invention
This invention relates to a fluorescence observation method of observing fluorescence emitted from fluorescent molecules like plural kinds of fluorescent proteins for example and relates to a florescence observation apparatus.
2. Description of Related Art
Observations of biomolecules using fluorescent molecules are standard methods of observation in the field of medicine or life science. Various fluorescent molecules which are different in fluorescence wavelength have been developed, and it becomes possible to observe plural kinds of biomolecules by using these fluorescent molecules in combination with one another.
In conventional methods of observing fluorescence emitted by a fluorescent molecule, for example, there exist: a method in which a fluorescent molecule is excited through one-photon excitation with light in the visible region and fluorescence generated on the longer-wavelength side of the wavelength of exciting light (Stokes shift) is detected; and a method in which a fluorescent molecule is excited through multi-photon excitation with light in the near-infrared region and fluorescence generated on the shorter-wavelength side of the wavelength of exciting light is detected (refer to Non Patent Literature 1, for example).
On the other hand, a method of exciting fluorescent proteins through one-photon excitation with light in the deep ultraviolet region is disclosed in the following Non Patent Literature 2. Non Patent Literature 2 discloses that fluorescent molecules (proteins) have absorption wavelength bands in the deep ultraviolet region and these fluorescent molecules have a common absorption wavelength band in the deep ultraviolet region even though these fluorescent molecules are different from one another.
As a result, if the fluorescent molecules are excited through one-photon excitation with light in the deep ultraviolet region and fluorescence generated on the longer-wavelength side of the wavelength of exciting light (Stokes shift) are detected, then it becomes possible to observe plural types of fluorescence simultaneously.